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. 1994 Aug;176(16):4838–4844. doi: 10.1128/jb.176.16.4838-4844.1994

Purification and characterization of a 1,2,4-trihydroxybenzene 1,2-dioxygenase from the basidiomycete Phanerochaete chrysosporium.

S Rieble 1, D K Joshi 1, M H Gold 1
PMCID: PMC196317  PMID: 8050996

Abstract

1,2,4-Trihydroxybenzene (THB) is an intermediate in the Phanerochaete chrysosporium degradation of vanillate and aromatic pollutants. A P. chrysosporium intracellular enzyme able to oxidatively cleave the aromatic ring of THB was purified by ammonium sulfate precipitation, hydrophobic and ion-exchange chromatographies, and native gel electrophoresis. The native protein has a molecular mass of 90 kDa and a subunit mass of 45 kDa. The enzyme catalyzes an intradiol cleavage of the substrate aromatic ring to produce maleylacetate. 18O2 incorporation studies demonstrate that molecular oxygen is a cosubstrate in the reaction. The enzyme exhibits high substrate specificity for THB; however, catechol cleavage occurs at approximately 20% of the optimal rate. THB dioxygenase catalyzes a key step in the degradation pathway of vanillate, an intermediate in lignin degradation. Maleylacetate, the product of THB cleavage, is reduced to beta-ketoadipate by an NADPH-requiring enzyme present in partially purified extracts.

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Selected References

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  1. Alic M., Letzring C., Gold M. H. Mating System and Basidiospore Formation in the Lignin-Degrading Basidiomycete Phanerochaete chrysosporium. Appl Environ Microbiol. 1987 Jul;53(7):1464–1469. doi: 10.1128/aem.53.7.1464-1469.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  3. Buswell J. A., Hamp S., Eriksson K. E. Intracellular quinone reduction in Sporotrichum pulverulentum by a NAD(P)H:quinone oxidoreductase: possible role in vanillic acid catabolism. FEBS Lett. 1979 Dec 1;108(1):229–232. doi: 10.1016/0014-5793(79)81216-8. [DOI] [PubMed] [Google Scholar]
  4. Chapman P. J., Ribbons D. W. Metabolism of resorcinylic compounds by bacteria: alternative pathways for resorcinol catabolism in Pseudomonas putida. J Bacteriol. 1976 Mar;125(3):985–998. doi: 10.1128/jb.125.3.985-998.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chapman P. J., Ribbons D. W. Metabolism of resorcinylic compounds by bacteria: orcinol pathway in Pseudomonas putida. J Bacteriol. 1976 Mar;125(3):975–984. doi: 10.1128/jb.125.3.975-984.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  7. Gold M. H., Alic M. Molecular biology of the lignin-degrading basidiomycete Phanerochaete chrysosporium. Microbiol Rev. 1993 Sep;57(3):605–622. doi: 10.1128/mr.57.3.605-622.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gold M. H., Kuwahara M., Chiu A. A., Glenn J. K. Purification and characterization of an extracellular H2O2-requiring diarylpropane oxygenase from the white rot basidiomycete, Phanerochaete chrysosporium. Arch Biochem Biophys. 1984 Nov 1;234(2):353–362. doi: 10.1016/0003-9861(84)90280-7. [DOI] [PubMed] [Google Scholar]
  9. Joshi D. K., Gold M. H. Degradation of 2,4,5-trichlorophenol by the lignin-degrading basidiomycete Phanerochaete chrysosporium. Appl Environ Microbiol. 1993 Jun;59(6):1779–1785. doi: 10.1128/aem.59.6.1779-1785.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kirk T. K., Farrell R. L. Enzymatic "combustion": the microbial degradation of lignin. Annu Rev Microbiol. 1987;41:465–505. doi: 10.1146/annurev.mi.41.100187.002341. [DOI] [PubMed] [Google Scholar]
  11. Kirk T. K., Lorenz L. F. Methoxyhydroquinone, an intermediate of vanillate catabolism by Polyporus dichrous. Appl Microbiol. 1973 Aug;26(2):173–175. doi: 10.1128/am.26.2.173-175.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  13. Spadaro J. T., Gold M. H., Renganathan V. Degradation of azo dyes by the lignin-degrading fungus Phanerochaete chrysosporium. Appl Environ Microbiol. 1992 Aug;58(8):2397–2401. doi: 10.1128/aem.58.8.2397-2401.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Trivedi L., Gupta A., Ki Paik W., Kim S. Purification and properties of protein methylase II from wheat germ. Eur J Biochem. 1982 Nov 15;128(2-3):349–354. doi: 10.1111/j.1432-1033.1982.tb06971.x. [DOI] [PubMed] [Google Scholar]
  15. Tuor U., Wariishi H., Schoemaker H. E., Gold M. H. Oxidation of phenolic arylglycerol beta-aryl ether lignin model compounds by manganese peroxidase from Phanerochaete chrysosporium: oxidative cleavage of an alpha-carbonyl model compound. Biochemistry. 1992 Jun 2;31(21):4986–4995. doi: 10.1021/bi00136a011. [DOI] [PubMed] [Google Scholar]
  16. Valli K., Brock B. J., Joshi D. K., Gold M. H. Degradation of 2,4-dinitrotoluene by the lignin-degrading fungus Phanerochaete chrysosporium. Appl Environ Microbiol. 1992 Jan;58(1):221–228. doi: 10.1128/aem.58.1.221-228.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Valli K., Gold M. H. Degradation of 2,4-dichlorophenol by the lignin-degrading fungus Phanerochaete chrysosporium. J Bacteriol. 1991 Jan;173(1):345–352. doi: 10.1128/jb.173.1.345-352.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Valli K., Wariishi H., Gold M. H. Degradation of 2,7-dichlorodibenzo-p-dioxin by the lignin-degrading basidiomycete Phanerochaete chrysosporium. J Bacteriol. 1992 Apr;174(7):2131–2137. doi: 10.1128/jb.174.7.2131-2137.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Wariishi H., Valli K., Gold M. H. In vitro depolymerization of lignin by manganese peroxidase of Phanerochaete chrysosporium. Biochem Biophys Res Commun. 1991 Apr 15;176(1):269–275. doi: 10.1016/0006-291x(91)90919-x. [DOI] [PubMed] [Google Scholar]
  20. Wojtaś-Wasilewska M., Trojanowski J., Luterek J. Aromatic ring cleavage of protocatechuic acid by the white-rot fungus Pleurotus ostreatus. Acta Biochim Pol. 1983;30(3-4):291–302. [PubMed] [Google Scholar]
  21. Wojtaś-Wasilewska M., Trojanowski J. Purification and properties of protocatechuate 3,4-dioxygenase from Chaetomium piluliferum induced with p-hydroxybenzoic acid. Acta Biochim Pol. 1980;27(1):21–34. [PubMed] [Google Scholar]

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